JP2017026621A - Detector and detection method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a simple detector capable of detecting a subject material in a small amount of a sample.SOLUTION: A detector for capturing a subject material in a sample at a prescribed position for detecting the same, comprises a detection tank for storing a liquid which may serve as a moving phase. The detection tank comprises in this order, a storage part which can store a liquid, a put-in part in which the sample is put, a detection part for detecting the subject material, and a liquid absorption part which can store a liquid absorption material for moving the liquid from the put-in part side to the detection part side, and a development member which can move the liquid and the sample therein while holding the same, is arranged so as to contact both of the storage part and the liquid absorption part. A channel disposed on the development member is configured so that a width or a cross section is reduced as going from a direction of a position 15 to a direction of a position 17 on any position in a region from the position 15 corresponding to the put-in part to the position 17 corresponding to the detection part, and the position 17 has a gap part where the development member does not exist.SELECTED DRAWING: Figure 4

Description

  The present invention relates to a detection apparatus and a detection method for detecting a test substance in a specimen.

There are various methods for detecting a test substance in a sample. Among them, a method is known in which a liquid and a sample are moved and a test substance in the sample is specifically detected by a detection unit. A typical example of such a detection method is a chromatography method. Chromatography is a method in which a specimen placed at one end of a stationary phase is moved with an appropriate mobile phase (liquid), and the difference in moving speed based on the difference in adsorption and distribution coefficient of each component in the specimen is used. Inspection / detection technology that separates
Patent Document 1 describes a high-performance liquid chromatography apparatus in which a means for moving a liquid and a specimen is a liquid delivery using a pump.
In particular, when a test substance in a biological sample is detected, an immunochromatography method has been used (for example, Patent Documents 2 and 3). In the conventional immunochromatography methods as described in Patent Documents 2 and 3, the liquid and the specimen are moved by utilizing the capillary action of the membrane that is a stationary phase. Patent Documents 2 and 3 describe that after the liquid moves to the end of the membrane by capillary action, the liquid is absorbed by the water absorption pad to suppress the back flow of the liquid.

JP 2002-267650 A JP 2001-059844 A JP 2005-156346 A

A conventional immunochromatography apparatus has a simple structure and can easily detect a test substance. However, since the conventional immunochromatography apparatus uses the capillary action of the membrane as described above, the moving speed of the liquid as the mobile phase and the moving speed of the specimen cannot be adjusted. When the affinity of the antibody is low, there is a problem that a sufficient antibody-antigen reaction time cannot be secured.
In addition, an immunochromatography apparatus having a simple structure that uses a liquid contained in a specimen as a mobile phase is known. Such an immunochromatography apparatus does not require an additional operation of adding a liquid as a mobile phase, and thus the operation is simple. However, if the amount of the sample is small, the amount of the liquid that is the mobile phase is insufficient, so that the movement of the sample due to capillary action stops halfway, and the sample does not reach the detection unit. For this reason, such an immunochromatography apparatus is easy to operate but requires a large amount of specimen.
In addition, in order to suppress the background in detection and increase the detection sensitivity and accuracy, there is a problem that it is necessary to separately add a washing solution for washing the immune complex in the detection unit. When a large amount of cleaning liquid is added to perform sufficient cleaning, the water retention force of the water absorbing pad as described above cannot suppress the back flow of the liquid, and there is a problem that detection accuracy is lowered. .
On the other hand, in the high performance liquid chromatography apparatus described in Patent Document 1, since the liquid is moved by a pump, problems such as liquid backflow do not occur. However, the high performance liquid chromatography apparatus requires a pump as described above, and cannot be said to be a simple apparatus.

Accordingly, an object of the present invention is to provide a simple detection apparatus that can detect a test substance in a small amount of a sample. And the detection apparatus which enables the detection of the quantitative test substance with a low background is provided.
Another object of the present invention is to provide a detection device capable of highly accurate detection. In a more preferred embodiment, it is also an object of the present invention to provide a detection device that can also quantitatively measure a test substance. In a further preferred embodiment, it is an object of the present invention to provide a detection device that does not cause a back flow of liquid.
The present invention also provides a detection device capable of increasing the cleaning power by increasing the speed at which the liquid passes through the position corresponding to the detection unit, and increasing the detection sensitivity and accuracy by suppressing the background in the specimen. The purpose is to do.
Another object of the present invention is to provide a detection device with improved detection sensitivity, accuracy, etc. by adopting a structure in which members that interfere with detection using light are removed in the region of the detection unit. To do.

  In order to solve the above problems, the present invention (1) is a detection device that detects a test substance in a specimen by capturing it at a predetermined position, and includes a detection tank that contains a liquid that can be a mobile phase. In the tank, a storage unit that can store the liquid, an input unit that inputs the specimen, a detection unit that detects the test substance, and the liquid is moved from the input unit side to the detection unit side. A liquid-absorbing part capable of accommodating the liquid-absorbing material, and a deploying member that can be moved while holding the liquid and the specimen inside is provided with the storage part and the liquid-absorbing part. Is arranged so as to be in contact with both, and the development member is provided with a flow channel capable of moving the liquid, and the flow channel is formed from the first position corresponding to the charging portion. It reaches the second position corresponding to the detection unit In any of the regions, the width or the cross-sectional area decreases as the first position moves from the first position to the second position, and the gap portion does not include the deployment member at the second position. There is provided a detecting device characterized by that.

According to the present invention, the liquid flow is formed by the liquid absorbing force of the liquid absorbing material in the detection tank by bringing the liquid absorbing material into contact with the liquid in the liquid absorbing portion. In this case, for the liquid absorption part, for example, a structure that allows the liquid absorption material to be charged in a detachable state, or a lid that can be opened and closed to have an opening in the top plate or wall of the liquid absorption part, etc. Or an opening that can be closed by a seal, a cover, or the like, and a structure in which the liquid absorbing material can be inserted from the outside through the opening can be employed.
Then, the test substance in the sample input from the input unit moves in the detection tank on the liquid flow formed by the liquid absorbing material or other liquid absorbing means, and is captured by the detection unit. Since the liquid continues to move even after the test substance is captured by the detection unit, substances other than the test substance can be prevented from being non-specifically captured by the detection unit. Therefore, according to the present invention, it is possible to provide a detection apparatus that enables detection of a quantitative test substance with low background.
Further, unlike the conventional immunochromatography apparatus, the capillary action of the membrane is not used, so that the detection apparatus of the present invention can detect the test substance in a small amount of sample.

Further, in the flow path provided in the developing member of the present invention, as described above, a liquid flow is generated by bringing the liquid-absorbing material into contact with the liquid in the detection tank, and the liquid is generated in the sample introduced from the input unit. The contained test substance moves along the surface or inside of the deployment member. Then, in this case, somewhere in the region from the first position corresponding to the input unit (in the vicinity of the input unit) to the second position corresponding to the detection unit (in the vicinity of the detection unit) It is more preferable that the width or the cross-sectional area of the flow path is reduced as it progresses (as it progresses) from the position of 1 to the direction of the second position, that is, the direction in which the liquid flows. Adopted. Therefore, the speed of the liquid flow at a position near the detection unit is increased, so that the cleaning power of the specimen (test substance) that is adsorbed on a carrier or the like is increased. In addition, the relative amount of the test substance with respect to the carrier or the like increases. As a result, the background in the specimen can be suppressed and the detection sensitivity and accuracy can be increased.
Further, the flow path provided in the deployment member in the present invention is based on the flow direction of the liquid, from an upstream position upstream of the first position to a downstream side of the second position. It is more preferable to adopt a configuration in which the width or the cross-sectional area decreases as the liquid flows in the flow direction until reaching the downstream position. In addition, from the upstream position upstream of the first position to the second position, or from the first position to the downstream position downstream of the second position. The width or the cross-sectional area may decrease as the liquid flows.
By adopting such a configuration, the region having the configuration is more extensive in relation to the range of the deployment member (or the flow path provided in the sample) used for measurement of the specimen (test substance). By providing a part of it for measurement, it is possible to generate a more stable liquid flow as a whole, to further increase the speed of the liquid flow, and to increase the detergency of the specimen (test substance). It can be raised further. As a result, the background in the specimen can be further suppressed and the detection sensitivity and accuracy can be increased.

In the detection apparatus of the present invention, the liquid does not move in the detection tank until the liquid absorbing material is brought into contact with the liquid by the operation unit. Therefore, a usage mode using an operation device that can simultaneously operate the operation units of a plurality of detection devices is also possible. That is, a plurality of detection devices are prepared, and a specimen is introduced into each. Then, the detection device into which the sample is introduced is installed in the operation device. Next, by operating this operation device, the operation units of the plurality of detection devices are operated simultaneously. Then, the movement of the liquid in the detection tank of a some detection apparatus can be started simultaneously. By using such a use mode, it is possible to prevent an error in the movement time of the liquid between the detection devices when a plurality of detection devices of the present invention are used.
In this case, it is preferable that the operating device is provided with reading means for reading the detection unit of the detection device of the present invention. By adopting such a configuration for the operating device, a plurality of detection devices of the present invention can be used to simultaneously detect test substances from a plurality of specimens.
The material used for the deployment member generally has the property of absorbing the light used for detection, and thus hinders detection using light. Therefore, the detection apparatus of the present invention can improve the light transmittance, detection sensitivity, accuracy, etc. at the time of detection in the detection unit by adopting a structure (gap part) from which the developing member having such properties is removed.

In addition, another preferable aspect (2) of the present invention is that the liquid absorbing portion includes a liquid absorbing material, a storage portion for storing the liquid absorbing material in a non-contact state with the liquid, and the liquid absorbing material as the liquid. The detection device according to (1), further including an operation unit that is brought into contact with the detection device.
In this case, by bringing the liquid-absorbing material into contact with the liquid in the liquid-absorbing portion, the liquid flow is formed in the detection tank by the liquid-absorbing force of the liquid-absorbing material.
Further, as described above, the liquid absorption part in the detection device of the present invention preferably includes a liquid absorption material, a storage part, and an operation part. By adopting such a configuration, the mode of the liquid absorption part is switched by the operation unit from the state where the liquid absorption material is stored in the storage part to the state where the liquid absorption material contacts the liquid in the detection tank. Can do.
Furthermore, in a preferred embodiment of the present invention, the storage portion is disposed above the detection tank so that a liquid absorbing material can be introduced into the detection tank, and the operation portion is configured such that the liquid is the liquid absorbing material. It is characterized by comprising a flow direction forming means for forming a liquid flow so as to face the direction.
By setting it as such a form, a liquid absorption material can be easily injected | thrown-in from a storage part to a detection tank. That is, it is possible to provide a simple structure in which the liquid flow is formed by dropping the liquid absorbing material from the storage portion by the operation portion and bringing it into contact with the liquid.
Furthermore, in a preferred embodiment of the present invention, the storage unit includes an isolation member that is disposed in the detection tank and isolates the liquid absorbing material from the liquid, and the operation unit includes the liquid in the liquid absorbing material. Flow direction forming means for forming a flow of liquid to face is provided.
Thus, by providing the storage part with the separating member that separates the liquid absorbing material from the liquid, the liquid absorbing material can be prevented from coming into contact with the liquid in a state where the liquid absorbing material is stored in the storage part. . That is, the flow of the liquid toward the liquid absorbing material is not formed until the operation unit is operated.

Furthermore, in a preferred embodiment of the present invention, at least a part of the storage part is made of a sheet-like material, and the operation part includes a sheet breaking means for breaking the sheet-like material.
By setting it as such a structure, a liquid-absorbing material can be thrown into the liquid in a detection tank by tearing a sheet-like material with a sheet | seat fracture | rupture means. That is, the act of breaking the sheet material in the storage unit starts the movement of the liquid and the specimen in the detection tank.

Furthermore, in a preferred embodiment of the present invention, the storage portion has a pressing surface on the opposite surface provided with the sheet-like material, and the sheet breaking means presses the pressing surface so that the sheet on the opposite surface is provided. It is characterized by breaking the material.
By setting it as such a structure, a sheet-like material can be broken by simple operation of pushing in a pushing surface.

Furthermore, in a preferred embodiment of the present invention, the pushing surface of the storage portion is made of a material that is deformed by stress.
If it becomes such a form, it will become easy to push in a pushing surface, and a sheet-like material can be broken easily.

Furthermore, in a preferred embodiment of the present invention, the pushing surface of the storage portion is movable in the vertical direction.
If it becomes such a form, it will become easy to push in a pushing surface, and a sheet-like material can be broken easily.

Furthermore, in a preferred embodiment of the present invention, at least one needle or blade is provided inside the pushing surface of the storage portion with the needle tip or blade edge facing the sheet-like material.
By setting it as such a form, the fracture | rupture of a sheet-like material becomes very easy.
In the present invention, it is also preferable that a tear tape is provided on the sheet material and the sheet material is broken by pulling the tear tape.
By adopting such a form, the sheet-like material can be broken by a simple operation of pulling the tear tape.

Moreover, another preferable aspect (3) of the present invention is characterized in that the detection tank includes a top plate portion covering an upper surface thereof, and the input portion is an input port provided in the top plate portion. The detection apparatus according to (1) or (2).
Thus, by providing a top plate part in a detection tank, it can prevent that the liquid in a detection tank spills.

Moreover, another preferable aspect (4) of the present invention is the detection device according to any one of (1) to (3), wherein the deployment member is wet with the liquid. is there.
By adopting such a configuration, it is possible to suppress the diffusion of the sample in the liquid in the detection tank when the sample is loaded from the loading unit.
In such a form of the detection apparatus, the specimen and the liquid move through the inside of the developing member and move in the detection tank toward the liquid absorbing material.

Another preferred embodiment (5) of the present invention is characterized in that the deployment member branches into at least two flow paths, and the flow paths are joined again. 4) The detection device according to any one of 4).
By branching the flow path in this way, it is possible to load a plurality of samples into the detection device at a time and simultaneously detect the test substances in each sample.

In addition, another preferred aspect (6) of the present invention includes a first specific binding substance that specifically binds to the test substance, and the first specific binding substance is located at a predetermined position of the detection unit. According to any one of (1) to (5), the test substance in the sample can be captured at a predetermined position by being fixed. This is a detection device.
The detection device of the present invention having such a configuration is very excellent in accuracy.

Another preferred embodiment (7) of the present invention comprises a second specific binding substance that specifically binds to the test substance or a complex of the test substance and the first specific binding substance, The detection apparatus according to (6), wherein the test substance, the first specific binding substance, and the second specific binding substance are configured to form a complex. .
By adopting such a form, the test substance captured by the first specific binding substance can be detected with high accuracy.
In a preferred embodiment of the present invention, a marker is added to or incorporated in the second specific binding substance.
By setting it as such a form, the detection of a test substance becomes very easy.

Furthermore, in a preferred embodiment of the present invention, the first specific binding substance and the second specific binding substance are antibodies.
Since an antibody is extremely excellent in specific binding ability, the detection device of the present invention having such a form has very high accuracy.

Another preferred embodiment (8) of the present invention is the detection device according to any one of (1) to (7), wherein the liquid-absorbing material is a water-absorbing polymer. is there.
Since the water-absorbing polymer can absorb and retain water several times its own mass, in the detection device of the present invention of this form, the movement of liquid in the detection tank is interrupted or reversely flows. There is no.

Moreover, this invention (9) uses the detection apparatus as described in any one of said (1)-(8), moves a liquid and a test substance by the liquid absorption force of a liquid absorption material, The present invention also relates to a test substance detection method including a step of capturing a test substance at a predetermined position.
As a result, in the method for detecting a test substance, the liquid and the specimen are moved by the liquid-absorbing force of the liquid-absorbing material, and the test substance in the specimen is captured at a predetermined position. To the method for starting the movement of the liquid.
According to the present invention, the start stage of the movement of the liquid can be controlled by a simple operation of contacting the liquid absorbing material with the liquid. Further, since the liquid is moved by the liquid absorbing material, the movement of the liquid is not interrupted or backflowed. Further, according to the detection method of the present invention, the test substance in a small amount of sample can be detected as compared with the conventional immunochromatography method.

Furthermore, the present invention (10) also relates to a method for adjusting the moving speed of the liquid by adjusting the mass of the liquid-absorbing material and / or the liquid in the detection method according to the above-mentioned (9).
As a result, in the test substance detection method including the step of moving the liquid and the specimen by the liquid-absorbing force of the liquid-absorbing material and capturing the test substance in the specimen at a predetermined position, the mass of the liquid-absorbing material and / or the liquid It also relates to a method of adjusting the moving speed of the liquid by adjusting.
According to the present invention, the moving speed of the liquid can be adjusted very simply.
Moreover, according to this invention, the moving speed of the said liquid can be raised in steps by increasing the mass of the liquid absorbing material to be used in steps.

According to the present invention, a simple detection device capable of detecting a test substance in a small amount of a sample can be provided.
In a preferred embodiment of the present invention, a detection device capable of highly accurate detection can be provided. In a more preferred embodiment of the present invention, it is possible to provide a detection apparatus capable of quantitatively measuring a test substance. Furthermore, in a preferred embodiment of the present invention, a detection device that does not cause a backflow of liquid can be provided.
In addition, according to the present invention, the detection apparatus capable of increasing the cleaning power by increasing the speed at which the liquid passes through the position corresponding to the detection unit, and suppressing the background in the specimen and increasing the detection sensitivity and accuracy. Can be provided.
In addition, according to the present invention, a detection device with improved detection sensitivity, accuracy, and the like by adopting a structure in which a material (such as a developing member) that hinders detection using light is removed in the region of the detection unit. Can be provided.

(A) It is a perspective view of the detection apparatus of Example 1. FIG. (B) The exploded view seen from the side of the detection apparatus of Example 1. FIG. (A) The top view of the detection apparatus of Example 1. FIG. (B) A fiber sheet provided in the detection device of Example 1. (A) The exploded view seen from diagonally downward of the detection apparatus of Example 1. FIG. (B) AA line sectional view of a detecting device of Example 1. FIG. FIG. 4 is a cross-sectional view of the detection device according to the first embodiment taken along line BB. 4 is a schematic diagram showing an antibody-antigen reaction in the detection apparatus of Example 1. FIG. (A) The perspective view of a storage part in the detection apparatus of Example 2, (b) Sectional drawing when this storage part is installed in the detection apparatus. (A) Perspective view of the detection apparatus of Example 3, (b) An exploded perspective view. The figure showing the measurement result of influenza A protein using the detection apparatus of Example 1. FIG. The figure which image | photographed the movement of the fluorescence labeled antibody in the case of changing the quantity of the sodium polyacrylate of the detection apparatus of Example 1 using the fluorescence microscope. The figure showing the measurement result of the fluorescence intensity of a detection part at the time of changing the quantity of the sodium polyacrylate of the detection apparatus of Example 1. FIG.

<1> Detection Device <1-1> Detection Device and Sample Hereinafter, embodiments of the detection device of the present invention will be described in detail.
The present invention is a detection device that detects a test substance in a specimen by capturing it at a predetermined position, and includes a detection tank that contains a liquid that can be a mobile phase, and the liquid is stored in the detection tank A reservoir that can store the sample, a detector that detects the test substance, a detector that detects the test substance, and a liquid absorber that can move the liquid from the inlet to the detector. The liquid part is arranged in this order, and further, a deployment member that can be moved while holding the liquid and the sample inside is arranged so as to contact both the storage part and the liquid absorption part. And a flow path capable of moving the liquid is provided in the development member, and the flow path is changed from a first position corresponding to the input section to a second position corresponding to the detection section. In any of the areas up to A detection apparatus having a configuration in which a width or a cross-sectional area decreases as it proceeds from a position 1 to a position 2, and a gap portion that does not include the developing member is provided in the position 2. It is.
The specimen that can be examined by the detection apparatus of the present invention is not particularly limited as long as it can be dissolved or dispersed in a liquid, and examples thereof include biological specimens, mixtures containing organic / inorganic compounds, and aqueous solutions. Specifically, examples of biological specimens include blood, urine, saliva, stool, biological samples, extracts from food samples, and mixtures and aqueous solutions containing organic / inorganic compounds include dust samples, Examples include soil samples, seawater and river water. The detection apparatus of the present invention is particularly useful in the inspection of biological specimens that require simple, rapid and highly accurate inspection.
Hereinafter, the configuration of the detection apparatus of the present invention will be described. Note that these are for explaining one embodiment of the present invention, and the present invention is not limited to these.

<1-2> Detection Tank The detection tank in the present invention is not particularly limited as long as it is a container-like structure capable of storing a liquid. That is, it is only necessary to have at least a bottom surface and a side surface so that the liquid can be stored. In preferable embodiment of this invention, the said detection tank is provided with the top-plate part which covers the upper surface.
The material of the detection tank is not particularly limited as long as it can store liquid without leaking. From the viewpoint of industrial production, the detection tank is preferably made of a thermoplastic resin.
And as a detection tank in the present invention, more preferably, after the bottom plate portion and the middle frame portion are thermocompression-bonded (fixed), a deployment member is sandwiched between the middle frame portion and the top plate portion, and thermocompression bonding is performed. Can be configured. In addition, the detection tank can be configured by thermocompression bonding (fixing) with a deployment member sandwiched between the substrate tank and the top plate portion. By using the thermocompression bonding method as described above, a relatively strong structure can be obtained with a method that is simple to some extent.
Moreover, as a detection tank in the present invention, more preferably, a double-sided pressure-sensitive adhesive sheet or the like is used and a substrate tank (or a base plate part and a middle frame part may be thermocompression bonded) and a ceiling provided with a developing member. The plate portion can be fixed and configured. In addition, it is preferable to use the thing (for example, optical transparent adhesive sheet etc.) of a property which does not prevent optical detection as a double-sided adhesive sheet.
In addition, it is more preferable that the adhesive surface on the side where the specimen flows in the double-sided pressure-sensitive adhesive sheet or the like is subjected to a treatment for preventing nonspecific adsorption to the biosystem sample / specimen. In this case, the treatment may be performed directly on at least one pressure-sensitive adhesive surface such as a double-sided pressure-sensitive adhesive sheet, or another film having a pressure-sensitive adhesive surface to which the treatment is applied (for example, a bioassay film or the like). ) May be superposed on a normal double-sided PSA sheet or the like to form an equivalent configuration as a whole. In addition, it is preferable to use a non-specific adsorption-preventing film such as a bioassay film that does not interfere with optical detection.
In addition, by using the double-sided pressure-sensitive adhesive sheet in this way, it can be more easily fixed, and further, a function for preventing nonspecific adsorption of the sample can be easily added, and as a result, detection noise. Can be reduced to increase detection sensitivity.

<1-3> Input Unit In the present invention, the configuration of the input unit is not particularly limited as long as the sample can be input from the outside of the detection tank into the detection tank.
As described above, when the top plate portion is provided in the detection tank, it is preferable to provide a loading port in the top plate portion and use the loading port as the loading portion.

<1-4> Detection part The detection part in this invention will not be specifically limited if it is the structure installed in the tank of the position away from the injection | throwing-in part, and can detect a test substance. In the case where the detection tank is provided with a top plate as described above, an opening is provided in a portion of the top of the detection tank that is above the detection unit so that the detection unit can be easily observed. The top plate is preferably made of a transparent material.
The distance from the loading unit to the detection unit is not particularly limited, but it is preferable to set the distance so that there is no possibility that the sample is directly loaded into the detection unit when the sample is loaded from the loading unit.
The detection of the test substance in the detection unit is performed by capturing the test substance. The mechanism for capturing the test substance is preferably configured to specifically capture the test substance and not capture substances other than the test substance.

In a preferred embodiment of the detection apparatus of the present invention for specifically capturing a test substance, the detection apparatus includes a first specific binding substance that specifically binds to the test substance, and the first specific binding substance is detected by the detection unit. By fixing at a predetermined position, the test substance in the sample can be captured and detected at the predetermined position. By adopting such a configuration, the accuracy of the detection device of the present invention increases.
The following are preferable as the first specific binding substance. That is, when the test substance is a biological substance such as a protein, an antibody is preferable. In addition, when the test substance is a nucleic acid such as DNA or RNA having a specific sequence, DNA or RNA having a sequence complementary thereto may be mentioned. And when a test substance is organic / inorganic substance other than said chemical substance, it is preferable that it is a substance couple | bonded specifically with these substances which are test substances.

In a more preferred embodiment of the present invention, the test substance or the complex of the test substance and the first specific binding substance is further provided with a second specific binding substance that specifically binds to the test substance, the first test substance, The specific binding substance and the second specific binding substance can form a complex. By adopting such a configuration, the detection accuracy of the detection device is improved in each stage.
In this case, the second specific binding substance is not particularly limited as long as it has the specific binding ability described above, and an antibody is preferable. In addition, when the test substance and the first specific binding substance are DNA or RNA as described above, ethidium bromide or the like that specifically binds to the double helix structure of the nucleic acid as the second specific binding substance. Preferable examples include incalators.
Moreover, it is preferable that a marker is added to or incorporated in the second specific binding substance. By setting it as such a form, a test substance can be detected simply. The marker is not particularly limited as long as it can be added to the second specific binding substance, and includes a fluorescent molecule, colloidal gold, colored and fluorescent fine particles, a substance exhibiting coloration or luminescence reaction, and a radioisotope. Compound etc. are mentioned. Moreover, a radioisotope is illustrated as what can be inherent in a 2nd specific binding substance as a marker.
Considering safety, ease of detection, and quantitativeness, it is particularly preferable to use a fluorescent molecule as a marker. In this case, the fluorescence emitted from the fluorescent molecule can be quantitatively measured using a commercially available fluorescent reader.

In a particularly preferred form of the invention, the first specific binding substance and the second specific binding substance are antibodies. According to such an embodiment, the detection device of the present invention is extremely excellent in sensitivity and accuracy due to the excellent specific binding ability of the antibody.
An embodiment in which the first specific binding substance and the second specific binding substance are antibodies will be described.
As the first specific binding substance that binds specifically to the test substance (hereinafter referred to as the first antibody), a commercially available one may be used, or it may be prepared according to a conventional method. .
As a mode of fixing the first antibody at a predetermined position of the detection unit, the first antibody may be fixed at a predetermined position of a developing member to be described later according to a conventional method, or an insoluble solid carrier ( The first antibody may be fixed to sepharose beads or the like, and the solid carrier may be placed at a predetermined position.
The antibody that is the second specific binding substance (hereinafter referred to as the second antibody) is not particularly limited as long as it is specific to the test substance or the complex of the test substance and the first antibody, and is commercially available. May be used, or those prepared according to a conventional method may be used.
In particular, an antibody specific to the test substance is preferably used as the second antibody, and from the viewpoint of improving detection accuracy, the second antibody preferably recognizes an epitope different from the first antibody. .
Moreover, it is preferable that a gold colloid or a fluorescent molecule is added to the second antibody as a marker. Addition of these markers to the second antibody can be performed according to a conventional method. A commercially available fluorescent molecule can be used without limitation as the fluorescent molecule added to the second antibody.

<1-5> Liquid Absorbing Portion “Liquid absorbing” in the present invention is to absorb liquid, and the liquid absorbing portion is liquid from the inlet side to the detecting unit side by absorbing liquid in the detection tank. The liquid absorbing part is a liquid absorbing material, a storage part for storing the liquid absorbing material in a non-contact state with the liquid in the detection tank, and the liquid absorbing material in contact with the liquid. An operation unit is provided.
Hereinafter, the configuration of the liquid absorption part will be described.

<1-5-1> Storage part The storage part in this invention takes the structure which accommodates a liquid absorbing material in the non-contact state with the liquid in the said detection tank. The configuration of the storage unit is not particularly limited as long as the liquid absorbing material can be stored so as not to contact the liquid in the detection tank.

In a preferred embodiment of the present invention, the storage portion is disposed above the detection tank so that a liquid absorbing material can be introduced into the detection tank. By setting it as such a form, a liquid absorption material can be easily injected | thrown-in from a storage part to a detection tank.
In this case, a member that separates the liquid absorbing material from the liquid in the detection tank may be provided in the storage unit. In addition, without providing such a member, the liquid absorbing material may be held on the ceiling portion of the storage unit so that the liquid absorbing material and the liquid in the detection tank do not contact each other.

In addition to the above form, in a preferred embodiment of the present invention, the storage unit includes an isolation member that is disposed in the detection tank and isolates the liquid absorbing material from the liquid, and the operation unit includes: A flow direction forming unit configured to form a flow of the liquid toward the liquid absorbing material;
In such an embodiment, since the storage portion is provided in the detection tank in which the liquid is accumulated, the isolation member is made of a material that does not transmit the liquid in order to bring the liquid absorbing material and the liquid into a non-contact state. It will be necessary.

<1-5-2> Operation unit The operation unit is a portion used for an operation of bringing the liquid-absorbing material stored in the storage unit into contact with the liquid in the detection tank. Below, it illustrates about the form of an operation part.
(1) Operation part provided with sheet breaking means In a preferred embodiment of the present invention, at least a part of the storage part is made of sheet-like material, and the operation part comprises sheet breaking means for breaking the sheet-like material.
When providing a storage part in the upper part of a detection tank, it is preferable to comprise the lower surface of a storage part with a sheet-like material.
Moreover, when arrange | positioning a storage part in a detection tank, it is preferable to comprise at least one part of the isolation member which is contacting the liquid in a detection tank with a sheet-like material.
The material of the sheet-like material is not particularly limited as long as it is a material that can be broken by the sheet breaking means, and more preferably a material that does not transmit water. Examples of the material for the sheet-like material include plastic, cloth, paper, and metal. From the viewpoint of waterproofness, durability with time, and corrosion resistance, the material of the sheet-like material is preferably plastic or metal.

In an embodiment of the present invention comprising a sheet breaking means, the storage portion comprises a pushing surface on the opposite surface provided with the sheet-like material, and the sheet breaking means is formed on the opposite surface by pushing the pushing surface. It is preferable to be realized by breaking the sheet-like material.
In order to realize the sheet breaking means by pushing the pushing surface, it is preferable that the pushing surface of the storage portion is made of a material that is deformed by stress. The indented surface is preferably made of a material that is not broken by stress. Examples of such a material include rubber and plastic.

  Further, when the sheet-like material is broken by pushing the pushing surface, it is also preferable that the pushing surface of the storage portion is movable in the vertical direction. That is, it is also preferable that the storage portion has a push button switch-like structure.

  When the sheet-like material is broken by pushing the pushing surface, it is preferable to provide at least one needle or blade whose needle tip or blade edge faces the sheet-like material inside the pushing surface of the storage portion.

  Further, as a form for breaking the sheet-like material, a form in which a tear tape is provided on the sheet-like material and the sheet-like material is broken by pulling the tear tape is also preferable.

(2) Operation unit provided with means for extracting isolation member In the above-described form in which the storage unit is installed in the detection tank, the liquid in the detection tank and the liquid absorbing material are isolated by the isolation member. In the case of such an embodiment, it is also preferable that the liquid in the detection tank is brought into contact with the liquid absorbing material by pulling out the separating member from the detection device to the outside.

(3) Operation part provided with means for rupturing balloon-like storage part It is also preferable to adopt a form like a balloon that bursts when the storage part is punctured with a needle or the like. With such a configuration, by piercing the storage portion with a needle or the like, the storage portion can be ruptured and the liquid absorbent material inside can be easily brought into contact with the liquid in the detection tank.

(4) Operation part provided with rotating means of storage part The form of the storage part is provided so as to be in close contact with the outside of the internal cylinder part and the internal cylinder part provided with an opening part in part. It is also preferable that the external cylinder portion is provided. In this case, it is preferable that the storage unit is provided in the upper part of the detection tank with the opening of the outer cylinder facing downward and the opening of the inner cylinder facing upward. In the detection device having the storage portion in such a form, the liquid absorbing material stored in the storage portion is obtained by rotating the internal cylinder portion and aligning the opening portion of the internal cylinder portion with the opening portion of the external cylinder portion. , Dropped into the detection tank.

<1-5-3> Liquid-absorbing material The liquid-absorbing material in the present invention refers to a substance having a property of absorbing liquid or a member containing such a substance. Examples of the liquid absorbing material include filter paper, cotton, sponge, cloth, sponge, and water-absorbing polymer.
As the liquid absorbing material in the present invention, a water absorbing polymer is particularly preferable. The water-absorbing polymer means a polymer having water absorption and water retention. In the present invention, it is particularly preferable to use a super absorbent polymer (abbreviation: SAP) among the super absorbent polymers. A highly water-absorbing polymer is a polymer designed to have a particularly high water-absorbing property and moisture retention performance, and has the property of incorporating a large number of water molecules into a gel structure in the network structure formed by the polymer. Is. The superabsorbent polymer can absorb 100 to 1000 times its own weight, and has a property that it is difficult to separate even when pressure is applied. That is, in the detection device of the present invention that employs a highly water-absorbing polymer as the liquid-absorbing material, the liquid does not substantially flow backward in the detection tank.
Such high water-absorbing polymers include starch-based and cellulose-based by graft polymerization or carboxymethylation, polyacrylate, polyvinyl alcohol, polyacrylamide, polyoxyethylene, and sodium polyacrylate. These synthetic polymer polymers can be mentioned. Particularly preferred is a sodium polyacrylate polymer from the viewpoint of water absorption and water retention functions.

In the case of using sodium polyacrylate, which is a highly water-absorbing polymer, as the water-absorbing polymer in the present invention, any of powder, granule, sphere, fiber, and sheet can be used. .
Moreover, the sodium polyacrylate to be used can be selected according to the property of the specimen put into the detection apparatus of the present invention and the purpose of the test.
It is known that sodium polyacrylate produced by an aqueous solution polymerization method is indefinite, and that produced by a suspension polymerization method becomes a microsphere. And since the microspherical sodium polyacrylate has a large surface area, the water absorption speed is larger than that of the amorphous one. In addition, amorphous sodium polyacrylate can absorb more water than microspheres.
That is, even when the test substance is captured by the detection unit, the detection unit is washed with a large amount of liquid, and when it is desired to prevent non-specific capture of substances other than the test substance by the detection unit as much as possible, A highly water-absorbing type sodium polyacrylate prepared by a legal method can be used. In addition, when it is desired to increase the moving speed of the liquid in the detection tank and obtain a detection result at an early stage, a rapid water absorption type sodium polyacrylate manufactured by suspension polymerization can be used.

Moreover, in this invention, the moving speed of the liquid in a detection tank can be adjusted by adjusting the mass of a liquid and / or a liquid absorbing material. That is, if the mass of the liquid absorbing material is increased, the moving speed of the liquid in the detection tank can be increased. Conversely, if the mass of the liquid absorbing material is decreased, the moving speed of the liquid can be decreased.
In addition, if the mass of the liquid is increased, the moving speed of the liquid in the detection tank can be decreased. Conversely, if the mass of the liquid is decreased, the moving speed of the liquid can be increased.
Furthermore, in this invention, the moving speed of a liquid can be adjusted by adding a liquid absorbing material in steps. As a form of the detection apparatus of the present invention in which the liquid absorbing material can be added stepwise, for example, a form provided with a plurality of storage units can be mentioned. In such a form of the detection device of the present invention, the liquid moving speed can be adjusted stepwise by bringing the liquid absorbing material stored in the respective storage portions into contact with the liquid stepwise.

  Furthermore, when using sodium polyacrylate as the liquid-absorbing material in the present invention, the moving speed of the liquid in the detection tank can be adjusted by changing the type of sodium polyacrylate.

<1-6> Deployment member In a preferred embodiment of the present invention, a deployment member capable of moving while holding the liquid and the specimen inside is provided in the detection tank, and the deployment member is wet with the liquid. It is characterized by.
The material used for the deployment member generally has the property of absorbing the light used for detection, and thus hinders detection using light. Therefore, the flow path provided in the developing member is processed so as to remove the member (that is, provide a gap and a cut) at a position corresponding to and facing the detection portion so as not to interfere with light detection.
Here, the developing member refers to a member having a fibrous structure or a dense structure of particles and capable of moving while holding a liquid and a specimen inside. Specifically, from a stationary phase used in thin layer chromatography, such as a thin plate of silica gel, alumina, polyamide resin, etc. on a glass plate, or from cellulose, resin, glass used in immunochromatography And a non-woven sheet, a gel composition such as agarose gel, and the like. In addition, it cannot be overemphasized that the expansion | deployment member in this invention is not restricted to what was enumerated above.
In addition, the flow path is provided by processing the shape of the developing member. In the flow path, as described above, a liquid flow is generated by bringing the liquid-absorbing material into contact with the liquid in the detection tank. The test substance contained in the sample to be introduced moves along the surface or inside of the development member. In this case, in the flow path, the first position corresponding to the input portion (near the input portion, the position below the input (possible) region) to the second position corresponding to the detection portion (near the detection portion, detection (possible) ) In any (somewhere) area in the area between (the position under the area)), as the liquid flows in the direction from the first position to the second position, that is, in the direction in which the liquid flows (along the traveling direction). The shape is narrowed down so as to reduce the width or cross-sectional area of the flow path, more preferably, the shape is processed so as not to increase. More preferably, it may be processed into a substantially rectangular shape from the position where the width or cross-sectional area size is narrowed to a certain extent until it reaches the detection portion from the position corresponding to the input portion. Good. It should be noted that a thin shape such as cloth or filter paper is preferable, but a three-dimensional thickness may be provided so as not to adversely affect analysis and detection.
Further, the middle frame portion or the substrate tank may be provided with a flow channel groove for accommodating the developing member.

Here, an embodiment in the case where a sheet-like member such as a nitrocellulose membrane or a fiber sheet is used as the developing member of the present invention will be described in detail.
The deployment member is provided with a gap that does not include the deployment member at a position corresponding to the detection unit in the flow path. The gap portion generally has a property of absorbing light used for detection by the developing member. Therefore, by removing the developing member having such a property, light transmittance, detection sensitivity, and the like in the detection unit are increased. Even if there is a gap (cut) in the developing member in which the flow path is disposed, the gap is filled in the liquid or the liquid is present, and the gap is kept at a distance within an appropriate range. If it is filled with a material having the property of holding a liquid, such as a bead-like carrier, so as to fill the gap, the flow of the liquid serving as the mobile phase and the accompanying specimen (test substance) ) Is not hindered.
Moreover, it is preferable that the expansion | deployment member in a detection tank is wet with the liquid before throwing in a test substance.
Moreover, it is preferable that the liquid exists only in the expansion | deployment member from an injection | throwing-in part to a detection part. Therefore, it is preferable to provide a storage part that can store liquid at both ends of the detection tank, and to make the bottom surface of the detection tank from the input part to the detection part higher than the bottom surface of the storage part. By doing so, it is possible to create a state in which the liquid exists only in the developing member from the input unit to the detection unit while accumulating a large amount of liquid in the storage unit.

In addition, although the development member can be configured to be branched into two or more in one flow path, in a preferred embodiment of the present invention, the development member is branched into at least two flow paths, and the flow paths are merged again. The structure to be taken is taken.
By branching the flow path in this way, it is possible to load a plurality of samples into the detection device at a time and simultaneously detect the test substances in each sample. It is also possible to perform comparative detection together with the reference.
In this case, it is preferable that the liquid does not move between the developing members forming the respective flow paths (at least the portion from the input portion to the detection portion). Specifically, it is preferable that a convex barrier portion that separates the respective flow paths is provided in the detection tank. In the case of adopting such a form, it is preferable to further provide a structure in which an input portion is provided independently for each flow path. This is from the viewpoint of preventing contamination in the detection tank of a plurality of specimens.

  As described above, when the insoluble solid carrier (sepharose beads or the like) to which the first antibody is immobilized is placed on the detection unit, a gap is provided in a portion corresponding to the detection unit of the development member, It is also preferable that the solid carrier is placed in the gap.

<2> Use of the detection device of the present invention by the operation device In the detection device of the present invention, the liquid does not move in the detection tank until the liquid absorbing material is brought into contact with the liquid by the operation unit. Therefore, a usage mode using an operation device that can simultaneously operate the operation units of a plurality of detection devices is preferable.
That is, a plurality of detection devices are prepared, and a specimen is introduced into each. Then, the detection device into which the sample is introduced is installed in the operation device. Next, by operating this operation device, the operation units of the plurality of detection devices are operated simultaneously. Then, the movement of the liquid in the detection tank of a some detection apparatus can be started simultaneously. By using such a use mode, it is possible to prevent an error in the movement time of the liquid between the detection devices when a plurality of detection devices of the present invention are used.
In this case, it is preferable that the operating device is provided with reading means for reading the detection unit of the detection device of the present invention. By adopting such a configuration for the operating device, a plurality of detection devices of the present invention can be used to simultaneously detect test substances from a plurality of specimens.

When using the detection device of the present invention in which the sheet-like material is broken by pushing the pushing surface described in <1-5-2> (1) above, the operation device includes a plurality of the present inventions. It is preferable to provide a mechanism capable of pushing the pushing surface of the detection device.
As described in <1-4> above, when a fluorescent molecule is added as a marker to the second specific binding substance, it is preferable that the operating device includes a fluorescence reading mechanism as a reading unit.

<3> Test substance detection method The present invention provides a test substance detection method including a step of moving a liquid and a specimen by liquid absorption force of a liquid-absorbing material and capturing the test substance in the specimen at a predetermined position. The present invention also relates to a method of starting movement of the liquid by bringing a liquid absorbing material into contact with the liquid.
The liquid absorption force in the present invention is a force that absorbs liquid. In addition, the specimen, the test substance, the step of capturing the test substance at a predetermined position, and the liquid absorbing material in the present invention are the same as those described in the above item <1>.

<4> A method for adjusting the moving speed of a liquid The present invention relates to a method for detecting a test substance, which includes a step of moving a liquid and a specimen by a liquid absorption force of a liquid absorbing material and capturing the test substance in the specimen at a predetermined position. The method of adjusting the moving speed of the liquid by adjusting the mass of the liquid absorbing material and / or the liquid.
According to the present invention, if the mass of the liquid absorbing material is increased, the moving speed of the liquid in the detection tank can be increased. Conversely, if the mass of the liquid absorbing material is decreased, the moving speed of the liquid can be decreased. .
In addition, if the mass of the liquid is increased, the moving speed of the liquid in the detection tank can be decreased. Conversely, if the mass of the liquid is decreased, the moving speed of the liquid can be increased.
Embodiments of the specimen, the test substance, the step of capturing the test substance at a predetermined position, and the liquid absorbing material in the present invention are the same as those described in the item <1>.

<1> Example 1: Detection Device Provided with Operation Unit Provided with Sheet Breaking Unit (1) Production of Detection Device of Example 1 The detection tank 12 of the detection device 1 of Example 1 is made of an acrylic top plate 121 (length 52 mm, width 18 mm, height 0.5 mm), middle frame portion 122 (length 52 mm, width 18 mm, height 3 mm), and bottom plate portion 123 (length 52 mm, width 18 mm, height 0.5 mm) ( FIG. 1B and FIG. 3A).
The top plate part 121 is provided with one input port 15 and one detection port 16. Further, the top plate 121 is provided with a rectangular (vertical 20 mm, horizontal 12 mm) cutout 121 a so as to install the box-shaped storage unit 14 (FIGS. 1B and 3A). The cutout 121a is formed in a rectangular shape that is slightly smaller than the bottom surface of the storage portion 14. Furthermore, the convex part 12a is provided in the back side of the top-plate part 121 (FIG. 3 (a), (b)).
The middle frame portion 122 is provided with two rectangular (vertical 20 mm, horizontal 12 mm) cutouts 122 a so that the storage portion 13 can be formed (FIGS. 1B and 3A).
The detection device 1 of Example 1 was manufactured as follows. First, the bottom plate portion 123 and the middle frame portion 122 were thermocompression bonded. Then, a desired amount of the liquid 3 was added to the storage part 13 formed by the bottom plate part 123 and the middle frame part 122. And the fiber sheet 2 was pinched | interposed between the middle frame part 122 and the top-plate part 121, and it thermocompression-bonded. Finally, the storage part 14 was installed by thermocompression bonding, and the detection device of Example 1 was manufactured (FIGS. 1A and 1B).

The configuration of the detection device 1 of Example 1 manufactured as described above will be further described.
The detection device 1 of Example 1 manufactured as described above includes an acrylic detection tank 12 (length 52 mm, width 18 mm, height 4 mm) (FIGS. 1 to 4). The top plate portion 121 provided on the upper surface of the detection tank 12 is provided with an insertion port 15 and a detection port 17. Moreover, the storage part 13 (length 20mm, width 12mm, depth 3mm) by which the liquid was stored is provided in the both ends of the detection tank 12 (FIG. 4). As shown in FIG. 4, a storage unit 14 including a pushing surface 142 and a sheet-like material 141 is provided on the upper portion of the detection tank 12. The pushing surface 142 is made of a hard plastic film, and the sheet material 141 is made of an aluminum foil. And in the accommodating part 14, the sodium polyacrylate 4 which is a superabsorbent polymer is accommodated.
A fiber sheet 2 is provided as a developing member in the detection tank. The fiber sheet 2 is provided with a gap portion 22 at a portion that contacts the detection unit 16. Sepharose beads 162 carrying the first antibody 163 are placed in the gap 22 (FIG. 5A).

  In the detection apparatus 1 of the first embodiment, by applying a downward stress in the vertical direction to the pushing surface 142 (FIG. 4B), the sheet-like material 141 is broken, and the sodium polyacrylate 4 is placed in the detection tank. It can be dropped (FIG. 4C). The dropped sodium polyacrylate 4 absorbs the liquid in the detection tank, so that the liquid starts to move in the direction of the sodium polyacrylate 4 in the detection tank (FIG. 4C).

Next, the mechanism of detection of the test substance by the antibody-antigen reaction in the detection tank of Example 1 will be described. Separose beads 162 carrying the first antibody 163 are placed on the detection unit 16 (FIG. 5A).
In this case, the second antibody 164 labeled with a fluorescent molecule may be provided upstream from the detection unit 16 (FIG. 5A), or the second antibody 164 may be introduced simultaneously with the specimen. Anyway.
When the specimen is introduced from the introduction section and sodium polyacrylate 4 is dropped into the detection tank and the movement of the liquid starts, the test substance 5 and the second antibody 164 in the specimen form a complex along with the movement of the liquid. In the formed state, it moves toward the detection part (FIG. 5B). When the immune complex reaches the detection unit 16, the test substance 5 and the first antibody 163 are combined to form a ternary complex of the first antibody 163 -test substance 5 -the second antibody 164. It remains in the part 16 (FIG. 5C). The fluorescence emitted from the fluorescent molecules bound to the second antibody 164 captured by the detection unit 16 is measured by a fluorescence reader, thereby detecting the test substance contained in the specimen and quantifying the amount thereof. Can be measured automatically.

<2> Example 2: Detection Device Provided with Operation Unit Provided with Tier Tape The detection device 1 of Example 2 is a detection device provided with an operation unit provided with a tear tape 143. FIG. 6A is a perspective view of the storage portion 14 as viewed from the bottom. As illustrated in FIG. 6A, a tear tape 143 is provided on the sheet-like material 141 on the bottom surface of the detection device 1 according to the second embodiment. And it is comprised so that the excess edge part of the tear tape 143 may come out of the detection tank 12 as shown in FIG.6 (b).
In the detection apparatus of Example 2, by pulling the tear tape 143 that has come out from the detection tank 12, the sheet-like material 141 is broken, and the sodium polyacrylate 4 stored in the storage unit 14 is detected in the detection tank. It is thrown in.

<3> Example 3: Detection device in which substrate tank and top plate part are bonded with adhesive sheet (1) Production of detection device of Example 3 Detection vessel 12 'of detection device 1' of Example 3 is made of acrylic. The top plate 121 ′ (52 mm long, 18 mm wide, 0.5 mm high) and the substrate tank 124 ′ (52 mm long, 18 mm wide, 3 mm high) are the basic skeletons (FIGS. 7A and 7B). )). The substrate tank and the top plate can be obtained by cutting and processing the same material as the detection tank.
A circular (diameter 10 mm) cutout 121a 'is provided at one end of the top plate 121' so that a water-absorbing polymer can be introduced. In addition, a rectangular cutout 121b 'is provided at the other end of the top plate 121' so that a liquid can be injected into the reservoir 13 '. Further, the top plate 121 ′ is provided with a loading port 15 ′ for loading a sample (FIG. 7B).
The substrate tank 124 ′ is provided with a circular groove (diameter 10 mm, depth 0.3 mm) so that a water-absorbing polymer can be placed thereon. Further, the substrate tank 124 ′ is provided with a rectangular (tub-shaped) reservoir 13 ′ (vertical 10 mm, horizontal 10 mm, depth 2.3 mm). Furthermore, a channel groove 6 ′ (depth 0.3 mm) is provided for accommodating the fiber sheet 2 ′ and forming a channel. The width of the channel groove 6 ′ is 10 mm from the storage part to the charging part, and decreases from 10 mm to 3 mm toward the detection part in the charging part, and 3 mm downstream from the charging part until reaching the liquid absorption part.
The detection device 1 ′ of Example 3 was manufactured as follows.
The fiber sheet 2 ′ was cut into a shape that matched the circular groove for placing the flow channel groove 6 ′, the reservoir 13 ′, and the water-absorbing polymer in the substrate tank 124 ′, and was bonded with an adhesive. The transparent bioassay film 19 ′ (3M, polyolefin tape 9795R) having a pressure-sensitive adhesive surface on one side is peeled off from one side of the optical transparent pressure-sensitive adhesive sheet 18 ′ (Nitto Denko Corporation, LUCIACS (registered trademark) CS9862UA). Adhering to the non-adhesive surface, the liner on the other surface of the optical pressure-sensitive adhesive sheet 18 ′ was peeled off and adhered to the top plate 121 ′. The optical transparent adhesive sheet 18 ′ and the bioassay film 19 ′ have cutouts 181a ′ and 191a ′ corresponding to the cutout 121a ′ of the top plate portion 121 ′, and 181b ′ and cutout corresponding to the cutout 121b ′. Cutouts 181c 'and 191c' are provided corresponding to the inlet 15 '. The liner on the adhesive surface of the bioassay film 19 ′ adhered to the top plate portion 121 ′ was peeled off, and the top plate portion 121 ′ was adhered to the substrate tank 124 ′ to produce the detection device 1 ′ of Example 3 (FIG. 7 (a)).

The detection device 1 ′ of Example 3 manufactured as described above will be further described. The detection device 1 ′ of Example 3 manufactured as described above includes an acrylic detection tank 12 ′ (length 52 mm, width 18 mm, height 3.5 mm) (FIGS. 7A and 7B). . A top plate 121 ′ provided on the upper surface of the detection tank 12 ′ is provided with an inlet 15 ′. The detection tank 12 ′ has a circular groove (diameter 10 mm, depth 0.3 mm) on which one of the superabsorbent polymers can be placed at one end, and a reservoir in which liquid is stored at the other end. 13 '(length 10mm, width 10mm, depth 2.3mm) (Fig.7 (a)). Note that the liquid may be appropriately injected after the entire detection device is manufactured.
A fiber sheet 2 'is provided in the detection tank as a developing member. The fiber sheet 2 'is provided with a gap 22' at a portion corresponding to the detection unit 16 '. Sepharose beads 162 'carrying the first antibody 163' are placed in the gap 22 '.

<3> Detection of Test Substance Using Detection Device of Example 1 Next, the detection device of Example 1 was used to detect the test substance in the sample and measure the amount of the test substance.
(1) Preparation of preparative antibody-labeled beads and fluorescent-labeled antibodies Activated Thiol Sepharose 4B beads (diameter 90 μm, GE Healthcare) were combined with anti-influenza A monoclonal antibodies (clone M2110169, Fitzgerald) to prepare anti-influenza A antibody-labeled beads ( Production of Sepharose beads 162 carrying the first antibody 163). An anti-influenza A monoclonal antibody (clone M322221, Fitzgerald) was labeled with Alexa Fluor 647 C2 Maleimide (Thermo scientific) to produce a fluorescently labeled anti-insulin antibody (production of a second antibody 164 labeled with a fluorescent molecule).

(2) Quantitative measurement of influenza A protein using the detection apparatus of Example 1 Using the detection apparatus of Example 1, protein (insulin) can be easily obtained from a small amount of sample (25 μL) in a short time (20 minutes). Quantitative measurement was performed.
A detection apparatus of Example 1 was prepared in which 75 μL of TBS buffer was provided in each of the storage sections 13 at both ends in the detection tank, and 3 mg of sodium polyacrylate (AQUALIC CA, Nippon Shokubai) was stored in the storage section 14.
In addition, the sodium polyacrylate in this test example is an acrylic acid polymer partial sodium salt cross-linked product.
Influenza A protein (A / NEW Caledonia / 20/99 (N1H1)) 0, 1, 10, 100, 1000, 10000 ng / mL (diluted with PBS) 25 μL of 1 μL of fluorescently labeled antibody was added from the input part Thereafter, the pushing surface 142 of the storage unit 14 was pushed with a finger, the sheet material 141 was broken, and the sodium acrylate 4 was put into the storage unit 13 on the downstream side. The time when the sodium polyacrylate 4 was added was set to 0 minute, and after 15 minutes, an image of the detection part was obtained with a fluorescence microscope, and the gray value was measured with image analysis software (ImageJ, NIH).

  When the influenza A protein was quantified using the detection apparatus of Example 1, a small amount of sample (25 μL) can be easily quantified in a chip in a short time (20 minutes) with 1 to 100 ng / mL of protein. (FIG. 8). It was also shown that 1000 ng / mL and 10000 ng / mL protein can be detected with high intensity.

(4) Examination of change in flow rate by changing the amount of the water-absorbing polymer 75 μL of TBS buffer is provided in each of the reservoirs 13 at both ends in the detection tank, and (i) 0 mg, (ii) 1 mg, (iii) ) A detection device was prepared in which 3 mg and (iv) 50 mg of sodium polyacrylate 4 (AQUALIC CA, Nippon Shokubai) were housed. In addition, the sodium polyacrylate in this test example is an acrylic acid polymer partial sodium salt cross-linked product.
A mixture of 1 μL of the fluorescently labeled second antibody 164 mixed with 25 μL of PBS is injected from the inlet 15, and then the sodium acrylate 4 is introduced from the storage part 14 to the storage part 13 on the downstream side by pushing the pushing surface 142. did. The time when the sodium polyacrylate 4 was charged was set to 0 minute, and observation was performed with a fluorescence microscope until 15 minutes later to obtain an image.

When 1 mg of acrylic acid polymer partial sodium salt cross-linked product (Aquaric CA, Nippon Shokubai) is charged in the detection device 1 provided with 75 μL in each of the reservoirs 13 at both ends, the fluorescently labeled second antibody 164 is charged. Even after 15 minutes, the fluorescence intensity was almost the same as 0 mg, and 1 mg was found to have a slow flow rate and insufficient washing (FIGS. 9 and 10).
On the other hand, when 3 mg is added, most of the fluorescently labeled second antibody 164 reaches the detection part 1 minute after the injection, and then flows further downstream from the detection part, at a flow rate sufficient for washing. It was found that there was (FIGS. 9 and 10).

1, 1 ′ detection device 12, 12 ′ detection tank 12 a convex portion 121, 121 ′ top plate portion 122 middle frame portion 123 bottom plate portion 124 ′ substrate tank 13, 13 ′ storage portion 14 storage portion 141 sheet-like material 142 pushing surface 143 Tier tapes 15 and 15 ′ Input ports 16 and 16 ′ Detection unit 162 Sepharose beads 163 First antibody 164 Second antibody 17 Detection port 18 ′ Transparent optical adhesive sheet 19 ′ Bioassay film 2 and 2 ′ Fiber sheet 21 , 21 'channel 22, 22' gap 3 liquid 4 sodium polyacrylate 5 test substance 6 'channel groove

Claims (10)

A detection device for detecting a test substance in a sample by capturing it at a predetermined position,
It has a detection tank that contains a liquid that can be a mobile phase,
In the detection tank, a storage unit that can store the liquid, an input unit that inputs a specimen, a detection unit that detects the test substance, and the liquid from the input unit side to the detection unit side A liquid-absorbing part capable of accommodating the liquid-absorbing material to be moved, and a deployment member capable of moving while holding the liquid and the specimen inside. It is arranged so that it touches both over the part,
The expansion member is provided with a flow path capable of moving the liquid, and the flow path is a region from a first position corresponding to the input section to a second position corresponding to the detection section. In any of the above, the width or the cross-sectional area is reduced as it advances from the first position toward the second position, and the gap portion that does not include the deployment member is provided in the second position. Provided,
A detection device characterized by that.   The liquid-absorbing part includes a liquid-absorbing material, a storage part that stores the liquid-absorbing material in a non-contact state with the liquid, and an operation unit that brings the liquid-absorbing material into contact with the liquid. The detection device according to claim 1. The detection tank includes a top plate that covers the top surface thereof,
The detection device according to claim 1, wherein the input unit is an input port provided in the top plate unit.   The detection device according to claim 1, wherein the deployment member is wet with the liquid.   The detection device according to any one of claims 1 to 4, wherein the deployment member is configured to branch into at least two or more flow paths, and the flow paths merge again. Comprising a first specific binding substance that specifically binds to the test substance;
The first specific binding substance is fixed at a predetermined position of the detection unit, so that the test substance in the sample can be captured at the predetermined position. The detection device according to any one of claims 1 to 5.   A second specific binding substance that specifically binds to the test substance or a complex of the test substance and the first specific binding substance, the test substance, the first specific binding substance, and the The detection apparatus according to claim 6, wherein the detection apparatus is configured to be able to form a complex with the second specific binding substance.   The detection device according to claim 1, wherein the liquid absorbing material is a water absorbing polymer.   A test comprising a step of moving the liquid and the specimen by the liquid absorption force of the liquid absorbent material and capturing the test substance in the specimen at a predetermined position using the detection device according to any one of claims 1 to 8. Substance detection method.   The detection method according to claim 9, wherein the moving speed of the liquid is adjusted by adjusting a mass of the liquid absorbing material and / or the liquid.

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